小鼠骨髓多能成体祖细胞的分离、培养及体外诱导分化为胰岛素分泌细胞的研究

The culture and differentiation of adult bone marrow-derived pluripotential mesenchymal stem cells into insulin-producing cells

目的:探讨培养骨髓多能成体祖细胞的条件和生物学特性及其向胰岛素分泌细胞分化的可能性。方法:采用文献报道的条件培养液培养小鼠骨髓多能成体祖细胞,观察其细胞形态、生长情况、细胞表面标志及mR-NA水平,检测其oct-4基因表达,并以含胰高糖素多肽-1的无血清培养液诱导分化细胞,基因水平检测与胰岛素分泌细胞有关的基因表达。结果:小鼠骨髓多能成体祖细胞类圆形,细胞核大,胞质少;细胞增殖能力尚可;细胞表面CD13+、CD44-、CD45-、MHCⅡ-;有oct-4基因表达;诱导分化后,可见与胰岛素分泌细胞有关的基因表达。结论:成功培养小鼠骨髓多能成体祖细胞,具有与文献报道类似的生物学特征;有被诱导分化为胰岛素分泌细胞的可能性。

AIM： To study the culture and characteristics of mouse adult bone marrow- derived pluripotential mesenchymal stem cells and its potential to differentiate into insulin secretion cells. METHODS： Cells were plated on 60% DMEM - LG and 40% MCDB - 201 medium supplemented with 2% fetal calf serum and 10/xg/L PDGF - BB, 10μg/L EGF and 1 × 10^6 U/ L LIF. The proliferation rate, phenotype and oct - 4 mRNA were tested. After it was plated on serum - free medium DMEM/F12 with GLP - 1 and nicotinamide, the nkx2.2 ngn3, pdx - 1 and insulin 2 mRNA were tested. RESULTS： The cells were round with large nucleus and scant cytoplasma. They were CD13^＋, CD44^- , CD45^- and MHC Ⅱ^-. Oct- 4 mRNA were present. The nkx2.2 pdx - 1 and insulin 2 mRNA were presented in cells plated on the inducing medium at 14 days. CONCLUSION： The adult bone marrow- derived pluripotent stem cells were cultured and they has the possibilities to be induced into insulin - secreting cells.